Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing

Jacy Fang; Isabella Salinas; Sarah San Vicente; Caroline Zielinski; Moshe Sade-Feldman

doi:10.3791/66766

Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing

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05:58 min

August 16, 2024

DOI: 10.3791/66766-v

Jacy Fang*^1,2, Isabella Salinas*^1,2, Sarah San Vicente*^1,2, Caroline Zielinski*^1,2, Moshe Sade-Feldman^1,2,3

¹Krantz Family Center for Cancer Research, Department of Medicine,Massachusetts General Hospital, ²Broad Institute of MIT and Harvard, ³Department of Medicine,Harvard Medical School

Overview

This protocol outlines the procedure for rapidly dissociating human and mouse tumor samples for single-cell RNA sequencing. This technique is crucial for understanding the immune response against tumors and identifying predictors of therapy resistance.

Key Study Components

Area of Science

Neuroscience
Immunology
Single-cell analysis

Background

Understanding mechanisms of response or resistance to immunotherapies.
Importance of real-time processing and analysis of patient samples.
Development of single-cell RNA sequencing techniques.
Need to investigate cellular compartments within tumors.

Purpose of Study

To study the immune response against tumors.
To identify predictors of therapy success.
To overcome resistance to standard immunotherapies.

Methods Used

Single-cell RNA sequencing
Rapid dissociation of tumor samples
Real-time analysis of molecular features
Investigation of tumor ecosystems

Main Results

Insights into the immune response mechanisms.
Identification of key immunological determinants.
Enhanced understanding of tumor microenvironments.
Potential targets for overcoming therapy resistance.

Conclusions

Single-cell RNA sequencing is vital for tumor analysis.
Real-time processing aids in understanding complex tumors.
Future research can focus on identified predictors of therapy.

Frequently Asked Questions

What is single-cell RNA sequencing?

Single-cell RNA sequencing is a technique that allows for the analysis of gene expression at the individual cell level, providing insights into cellular heterogeneity.

Why is rapid dissociation of tumor samples important?

Rapid dissociation preserves the integrity of the cells and allows for timely analysis of their molecular features, which is crucial for understanding tumor biology.

How does this study contribute to immunotherapy research?

This study helps identify predictors of immune response and resistance, which can inform the development of more effective immunotherapy strategies.

What are the challenges in studying human immunity?

Challenges include the complexity of human immune responses and the need for real-time analysis of patient-derived samples.

What advancements have been made in tumor analysis?

Recent advancements include the integration of single-cell RNA sequencing techniques that allow for unbiased analysis of tumor ecosystems.

What is the significance of understanding tumor ecosystems?

Understanding tumor ecosystems is crucial for identifying how different cell types interact and contribute to tumor growth and response to therapies.

This protocol outlines the procedure for rapidly dissociating human and mouse tumor samples for single-cell RNA sequencing.

The scope of our research is to study the mechanisms of response or resistance to standard of care immunotherapies. Our lab is trying to understand what drives a successful immune response against tumors, and if we can identify these predictors of therapy to find targets that can overcome resistance. The biggest advancement in recent years has been the development and integration of single-cell RNA sequencing techniques.

They enable us in real time to systematically study molecular features in individual cells in an unbiased manner, which is helpful in the dissection of these complex and evolving tumor ecosystems. While many preclinical models are being used to study the response mechanisms to immunotherapy, we still don't know the immunological determinants that are important in human immunity. Processing and analyzing patient samples in real time is crucial to our ability to investigate all cellular compartments within a given tumor.

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